TABLE 2.
Synthetic oligonucleotide primers used for cloning and expression of mutated and truncated ORF2 proteins of the CaHEV strain and the N protein of the PRRSV SD16 strain
| Oligonucleotide | Sequenced | Motif |
|---|---|---|
| aORF2 355-570-Sa | CGGGATCCGTGAAATTGTATATGT | 355VKLYM |
| aORF2 360-570-S | CGGATCCTCGGTGGAGGATGCTGTC | 360SVEDAV |
| aORF2 365-570-S | CGGATCCGTCAACGACAAGCCCATT | 365VNDKPI |
| aORF2 371-570-S | CAAGGATCCATGGTCCCTCACGACATTGA | 371MVPHD |
| aORF2 380-570-S | GAAGGATCCACTAGCACTGTTACCTGC | 380TSTVTC |
| aORF2 PH373-374RR-Sb | AAAGGATCCATGGTCCGTCGCGACATTGAC | 371MVRRD |
| aORF2 P373R-S | AAAGGATCCATGGTCCGTCACGACATTGAC | 371MVRHD |
| aORF2 H374R-S | CAAGGATCCATGGTCCCTCGCGACATTGAC | 371MVPRD |
| aORF2 V372A-S | CAAGGATCCATGGCCCCTCACGACATT | 371MAPHD |
| aORF2 D375E-S | CAAGGATCCATGGTCCCTCACGAGATTGACCT | 371MVPHE |
| aORF2-Rc | GAACTCGAGGGTGGTGCAATCATCAC | |
| N Protein-S | CGGGATCCATGCCAAATAACAACGGCAAG | |
| N Protein-R | AACTCGAGTCATGCTGAGGGTGATGCTGT |
a
Forward primer encoding a truncated protein for epitope 1B5.
b
Forward primer encoding a recombinant protein (aa 371 to 570) for epitope 3E8, with aa P373 and H374 mutated to R.
c
Reverse primer for 10 mutation and truncation proteins from avian HEV ORF2.
d
Letters in italics indicate BamHI or XhoI sequences.